Abstract
Gap junctions are essential for the proper function of many native mammalian tissues including neurons, cardiomyocytes, embryonic tissues, and muscle. Assessing these channels is therefore fundamental to understanding disease pathophysiology, developing therapies for a multitude of acquired and genetic conditions, and providing novel approaches to drug delivery and cellular communication. Microinjection is a robust, albeit difficult, technique, which provides considerable information that is superior to many of the simpler techniques due to its ability to isolate cells, quantify kinetics, and allow cross-comparison of multiple cell lines. Despite its user-dependent nature, the strengths of the technique are considerable and with the advent of new, automation technologies may improve further. This text describes the basic technique of microinjection and briefly discusses modern automation advances that can improve the success rates of this technique.
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Acknowledgements
Special thanks to the following individuals for their hard work in helping to develop this protocol and the experiments for the testing thereof: Vinayakumar Siragam, Zheng Gong, Jun Chen, Clement Leung, Zhe Lu, Changhai Ru, Shaorong Xie, Jun Luo, and Lynn Strandberg. This work was supported by the Canadian Institutes of Health Research Team grant for ARVC Research (2009–2014), CIHR/NSERC Collaborative Health Research Project (2015–2018), The Caitlin Elizabeth Morris Fund of Appliance Canada, The Alex Corrance Memorial Fund, and the University of Toronto Paediatric Research and Clinical Summer Scholarship.
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Fridman, M.D., Liu, J., Sun, Y., Hamilton, R.M. (2016). Microinjection Technique for Assessment of Gap Junction Function. In: Vinken, M., Johnstone, S. (eds) Gap Junction Protocols. Methods in Molecular Biology, vol 1437. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3664-9_10
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DOI: https://doi.org/10.1007/978-1-4939-3664-9_10
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